System and method for in ovo sexing of avian embryos

ABSTRACT

A system for determining the gender and/or fertility status of avian eggs including a sampling apparatus and an electromagnetic radiation transmitter and detector. In certain embodiments, the transmitter operates in the terahertz range. The sampling apparatus can be coupled to an avian egg. The sampling apparatus includes a vacuum source, a gas collection device, and a membrane that can be positioned in the passageway coupling the vacuum source to the gas collection device. The membrane is capable of capturing volatile organic compounds. The sampling apparatus applies a vacuum from the vacuum source to the gas proximate to the avian egg and directs the gas captured from the vicinity of the egg toward the membrane. Subsequently, the membrane is positioned within the electromagnetic radiation emitted by the transmitter, generating a spectrum which can be analyzed to determine whether the egg is fertile or infertile, and if fertile, whether the egg is male or female. In an embodiment, the captured volatile organic compounds are

BACKGROUND OF THE INVENTION 1. Field of the Invention

The invention generally relates to the in ovo gender determination andfertility verification of avian embryos. More specifically, theinvention relates to rapid multiple egg in ovo gender and fertilitydetermination in a commercial setting.

2. Description of the Relevant Art

In commercial poultry production, one gender may be selected overanother for various practical and economic reasons. For the productionof table eggs, females are selected for their ability to lay eggs, andmales and infertile eggs are culled. In the case of broilers, males andfemales are often separated due to their different growth rates and timeto maturity, making it more economical to raise and feed themseparately.

In order to determine sex, trained human sexers are used to identify thebird's gender within a day of hatching, usually by feather length or viacloacal anatomy. Culling of the unwanted gender often takes place viamaceration or suffocation. The waste product from the culled chicks isthen disposed of, often at additional cost to the hatchery. Because thebirds must first be hatched in order to have their gender identified,hatcheries require twice the incubation and hatching capacity needed toproduce a given number of female laying hens (the sex ratio beingroughly 50:50 male to female) or male broilers.

While known art has been proposed for in ovo sexing, these techniquesare invasive, requiring puncturing the shell and membranes, to eitherremove a sample (e.g., of the blastoderm) or gain entry to directlyilluminate (in the case of spectrometric techniques) egg componentscontaining genetic material or other bio-identifying molecules andproteins. In addition to invasiveness, the aforementioned techniquesrely on measuring phenomena such as differences in the mass of geneticmaterial, feather coloring, sex-linked proteins, nucleotides, and thelike, or differences in sex hormones. In all cases these identifiersemerge only after several days of incubation, requiring hatcheries tomaintain enough capacity for these additional eggs. In a similar manner,using the aforementioned techniques, infertile eggs must be incubatedfor some time before lack of embryo development is evident.

Identifying a bird's gender in ovo would enable hatcheries to reduceincubation and hatchery operations and eliminate labor-related sexingcosts, resulting in gains in productivity and efficiency for the plant.The earlier eggs with relevant traits can be identified, the sooner theycan be segregated (e.g., in the case of broilers) or culled (e.g., inthe case of laying hens). This creates the maximum economic benefit tothe hatchery by limiting the amount of time eggs that are destined forseparation or culling must be incubated. The solutions mentioned in theprior art require at least some incubation before the egg can beproperly identified and handled. Thus, there is need in the market for asolution that can identify the gender or fertility of an eggnon-invasively and before an egg requires incubation.

SUMMARY OF THE INVENTION

In an embodiment, a system for determining the gender and/or fertilitystatus of avian eggs includes a sampling apparatus and anelectromagnetic radiation transmitter and detector. The transmitter mayoperate in, in various embodiments, the terahertz range, the microwaverange, or the infrared range. The sampling apparatus can be coupled toan avian egg. The sampling apparatus includes a vacuum source, a gascollection device, and a membrane that can be positioned in thepassageway coupling the vacuum source to the gas collection device. Themembrane is capable of capturing volatile organic compounds. Thesampling apparatus applies a vacuum from the vacuum source to the gasproximate to the avian egg and directs the gas captured from thevicinity of the egg toward the membrane. Subsequently, the membrane ispositioned within the electromagnetic radiation emitted by thetransmitter.

In an embodiment, a method of determining the gender and/or fertilitystatus of avian eggs includes: coupling a sampling apparatus to an avianegg, in which the sampling apparatus includes a vacuum source, a gascollection device, and a membrane capable of capturing volatile organiccompounds; operating the sampling apparatus to capture gas proximate tothe avian egg via the gas collection device, in which the samplingapparatus directs the captured air toward the membrane; and applyingelectromagnetic radiation to the membrane, and in which theelectromagnetic radiation is preselected to allow the quantity of one ormore volatile organic compounds captured in the membrane to bedetermined.

In an embodiment, a system for determining the gender and/or fertilitystatus of avian eggs includes a sampling apparatus and anelectromagnetic radiation transmitter and detector. The samplingapparatus applies a vacuum from the vacuum source to the gas proximateto the avian egg and directs the gas captured from the vicinity of theegg toward a sample chamber. The sample chamber is positioned within theelectromagnetic radiation emitted by the transmitter.

In an embodiment, a method of determining the gender and/or fertilitystatus of avian eggs includes: coupling the sampling apparatus to theavian egg; operating a sampling apparatus to capture gas proximate tothe avian egg via the gas collection device, in which the samplingapparatus directs the captured gas into the sample chamber; and applyingelectromagnetic radiation to the sample chamber, in which theelectromagnetic radiation is preselected to allow the quantity of one ormore volatile organic compounds captured in the sample chamber to bedetermined.

In an embodiment, a system for determining the gender and/or fertilitystatus of avian eggs includes one or more carriers for receiving one ormore avian eggs, one or more transmitters capable of emittingpreselected electromagnetic radiation, and one or more detectors. Thetransmitters direct the preselected electromagnetic radiation into theone or more avian eggs. As a non-exhaustive example, tunable continuouswave terahertz radiation (CW-THz) can be generated via photomixingtechniques known in the art using two Ti:sapphire lasers in adjacentwavelengths to generate a beat frequency in the desired range. Numerousantennae arrays are available for emitting the terahertz radiation,often consisting of GaAs structures. Many variations exist in theparticulars of creating terahertz (and other spectrum) radiation; asuitable device may be used that most efficiently produces the desiredwaveband. The detectors are capable of detecting the preselectedelectromagnetic radiation that passes through the one or more avian eggsusing proper antennae, electro-optical sampling methods, or othersuitable techniques.

In an embodiment, a method of determining the gender and/or fertilitystatus of avian eggs includes: positioning the air sac of an egg in theoptical pathway between a transmitter and a detector; and applyingelectromagnetic radiation to the air sac of the egg through the unbrokenshell, in which the electromagnetic radiation is preselected to allowthe quantity of one or more organic compounds in the air sac of the eggto be determined.

BRIEF DESCRIPTION OF THE DRAWINGS

Advantages of the present invention will become apparent to thoseskilled in the art with the benefit of the following detaileddescription of embodiments and upon reference to the accompanyingdrawings in which:

FIG. 1 illustrates an exemplary commercial vacuum egg handling system inone embodiment;

FIG. 2 depicts a schematic diagram of a sub-manifold of a vacuum egghandling system including a membrane to capture VOCs from eggs that aredisposed in egg handling cups;

FIG. 3 is a top view illustrating an exemplary system for analyzing andsorting eggs that includes dual reading trays and a movable readingdevice;

FIG. 4 is a front view illustrating an exemplary system for analyzingand sorting eggs that includes dual reading trays and a movable readingdevice;

FIG. 5 depicts a schematic diagram of a sub-manifold of a vacuum egghandling system that includes a sample chamber to capture VOCs from eggsthat are disposed in egg handling cups;

FIG. 6 depicts a schematic diagram of an exemplary in ovo detectionsystem whereby the air sac is in the bottom position, requiring thespectrometer to be placed beneath the eggs, and special trays allowingthe area of the egg containing the air sac to be exposed to thespectrometer;

FIG. 7 depicts a schematic diagram of a sub-manifold of a vacuum egghandling system that includes a single sample chamber to analyze VOCsfrom eggs that are disposed in egg handling cups; and

FIG. 8 depicts a schematic diagram of a sub-manifold of a vacuum egghandling system that includes a membrane sample chamber to capture VOCsfrom eggs that are disposed in egg handling cups.

While the invention may be susceptible to various modifications andalternative forms, including reversing the position of the vacuum to thebottom of the egg when the egg is oriented with the air sac facing down,e.g. using trays like that depicted in FIG. 6, specific embodimentsthereof are shown by way of example in the drawings and will herein bedescribed in detail. The drawings may not be to scale. It should beunderstood, however, that the drawings and detailed description theretoare not intended to limit the invention to the particular formdisclosed, but to the contrary, the intention is to cover allmodifications, equivalents, and alternatives falling within the spiritand scope of the present invention as defined by the appended claims.For example, though the drawings typically depict the egg with the airsac in the top position, if eggs are transported or moved about with theair sac in the bottom position, the invention allows for the apparatusto be reversed, with the vacuum collection taking place from the bottomof the egg (i.e., the location of the air sac in this example).

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

It is to be understood the present invention is not limited toparticular devices or methods, which may, of course, vary. It is also tobe understood that the terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to belimiting. As used in this specification and the appended claims, thesingular forms “a”, “an”, and “the” include singular and pluralreferents unless the content clearly dictates otherwise. Furthermore,the word “may” is used throughout this application in a permissive sense(i.e., having the potential to, being able to), not in a mandatory sense(i.e., must). The term “include,” and derivations thereof, mean“including, but not limited to.” The term “coupled” means directly orindirectly connected.

As used herein the term “avian eggs” refers to the eggs obtained fromavian species. Exemplary avian species include, but are not limited to,chickens, turkeys, ducks, quails, and geese.

Systems and methods disclosed herein addresses shortcomings of othermethods of in ovo gender and fertility determination by measuringvolatile organic compounds (VOCs) in the egg's air cell through theshell or in atmosphere (either directly or via a collecting membrane orvia a collecting chamber), enabling non-invasive detection of gender andfertility status prior to incubation. These volatiles vary throughoutthe incubation period, and have been shown to differ between fertile andinfertile eggs, as well as being predictive of embryo sex as early asday 1 of incubation. The type and quantity of typical VOCs emitted byavian eggs are discussed in the paper by Webster et al. “Avian Egg OdourEncodes Information on Embryo Sex, Fertility and Development” (2015)PLoS ONE 10(1): e0116345, and Costanzo et al, “The Odour of Sex:Sex-Related Differences in Volatile Compound Composition among BarnSwallow Eggs Carrying Embryos of Either Sex” (November 2016) PLOS ONEDOI:10.1371/journal.pone.0165055, both of which are incorporated hereinby reference.

Eggs diffuse volatile organic compounds, as well as CO₂, H₂O, and O₂,readily through their pores. Up to 20 L of oxygen, carbon dioxide andwater vapor can diffuse through the pores of an 80 gm egg by the timeinternal pipping takes place. These high vapor pressure volatiles can bedetected using spectroscopic techniques.

Membrane System

In one embodiment, a system for determining the gender and/or fertilitystatus of avian eggs includes a sampling apparatus that can be used tocollect volatile organic compounds (VOCs) from an intact egg. In oneembodiment, the sampling apparatus includes a vacuum source and amembrane capable of capturing volatile organic compounds. Any membranecapable of capturing VOCs of interest can be used. In one embodiment, apolyethylene terephthalate (PET) membrane is used. An exemplarypolyethylene terephthalate membrane is described in U.S. ProvisionalPatent Application No. 62/326,857 entitled “Avian Gender Classificationin Egg Prior to Hatch Using Chemical Indicators in THz spectrum”, filedApr. 25, 2016, which is incorporated herein by reference.

During use, a sampling apparatus applies a vacuum from the vacuum sourceto the gas proximate to the avian egg and directs the gas captured fromthe vicinity of the egg toward the membrane. In an embodiment, thesampling apparatus includes a gas collection device which is placedproximate to the egg. The gas collection device may have a number ofshapes chosen to optimize the collection of VOCs from a single egg. Inan embodiment, the gas collection device has a shape complementary tothe shape of a single egg (e.g., is “egg shaped” or ovoid). In oneembodiment, the gas collection device is the egg contact portion of avacuum egg lifter.

In one embodiment, off-gassed VOCs are adsorbed onto the membranes. The“loaded” membranes are then analyzed by applying electromagneticradiation (e.g., between 600-750 in the case of the terahertz part ofthe spectrum, though other bands of the electromagnetic spectrum may beused) to the membrane and observing the change in the electromagneticradiation. Analysis of the membrane may be accomplished using anelectromagnetic radiation transmitter and an electromagnetic radiationdetector typical of a spectrometer operating at microwave, terahertz orinfrared wavelengths. During analysis the membrane is positioned withinthe beam of electromagnetic radiation emitted by the transmitter. Theelectromagnetic radiation passes into the membrane and the interactionof the VOCs trapped in the membrane alter the electromagnetic radiation.After contacting the membrane, the altered electromagnetic radiation iscaptured by the electromagnetic radiation detector. The changes in theelectromagnetic radiation can be used to determine what VOCs are beingreleased by the egg. Male, female, fertile and infertile eggs allrelease a unique combination of VOCs. By analyzing the type and amountof VOCs the gender of the egg, and the status of an individual egg canbe determined.

Electromagnetic radiation in the microwave, terahertz or the infraredrange may be used to analyze VOCs. The analysis spectra may be generatedusing absorbance, transmittance, reflectance, or Raman spectroscopy.

In a preferred embodiment, terahertz electromagnetic radiation is usedfor the detection of VOCs captured in a membrane. As used hereinterahertz electromagnetic radiation refers to radiation having awavelength of between 1 mm to 0.01 mm. In a particular embodiment,terahertz radiation within the 600-750 μm range is used to determine theVOC content in a PET membrane. The electromagnetic radiation detectorgenerates an absorption spectrum. Absorption spectra can be obtained inthe frequency domain, or in the time domain and translated to frequencyvia Fourier transform, depending on the spectroscopic method used.

The absorption spectra is read and compared to a database via softwarematching algorithms. The database contains spectral fingerprints of eggswith each fingerprint representing fertility status or gender forvarious avian species and breeds within a species. The software matchingalgorithm compares the collected spectrum to the catalogued fingerprintwithin pre-determined confidence bounds, and identifies the gender andfertility status by determining whether or not the read spectrum fallswithin the error bounds of the fingerprint.

Once the status of the egg is determined, eggs of the culled sex arekicked off the line using conventional egg-sorting machinery and can beretained for sale as breaking stock in the case of eggs from layerbreeds and can be sorted by gender in the case of eggs from broilerbreeds among other purposes. In a similar manner, infertile eggs can beseparated from fertile eggs.

In an embodiment, membranes may be recycled via application ofelectricity to release the VOCs from the membrane. The “cleaned”membrane is cycled back into place on the sampling apparatus. In otherembodiments, the membrane can be cleaned by reversing the flow of thevacuum motor, which causes air to pass through the membrane and push theabsorbed molecules from the membrane. In certain embodiments, eachmembrane may be used only once and then replaced by a new membrane.

An exemplary commercial vacuum egg handling system loader (in this case,Moba FL330 loader) is depicted in FIG. 1 as an example ofvacuum-handling equipment in use by a large number of hatcheries. Adevice similar to that shown can be used to implement the invention in acommercial setting. Incorporating a sampling apparatus into a commercialvacuum egg handling system may allow more than 100 eggs to be sampledsimultaneously by the vacuum handler. Turning to FIG. 1, a vacuum egghandling system 100 generally includes a vacuum source 110 and aplurality of egg handling cups 120 coupled to the vacuum system. In anembodiment, the vacuum source 110 is coupled to the egg handling cups120 via one or more vacuum manifolds. In the particular embodimentdepicted in FIG. 1, vacuum source 110 is coupled to egg handling cups120 via first vacuum manifold 130 and a plurality of sub-manifolds 140.As shown, vacuum source 110 is coupled to first vacuum manifold 130 viavacuum source conduit 115. First vacuum manifold 130 is coupled to aplurality of sub-manifolds 140 via a plurality of manifold conduits 135.A plurality of egg handling cups 120 are coupled to each of thesub-manifolds. The vacuum created in vacuum source 110 is conducted toeach of the sub-manifolds creating a vacuum in the egg handling cup,sufficient to pull an egg into the egg handling cup and hold the egg inthe egg handling cup while the egg is being transported, as shown inFIG. 1.

In one embodiment, a vacuum egg handling system, such as shown in FIG.1, may be modified to include a plurality of sampling apparatuses. FIG.2 depicts a schematic diagram of a sub-manifold 240 of a vacuum egghandling system that is modified to include a membrane 250 to captureVOCs from eggs that are disposed in egg handling cups 220. In thisembodiment, egg handling cups 220 act as the gas collection device. Gasin the vicinity of the egg is pulled through the egg handling cups 220into the membrane 250 where VOCs that are produced by the egg arecollected. In some embodiments, membranes may be coupled to an electricsystem to impart a small charge to the membrane to help withadsorption/desorption of VOCs.

As depicted in FIG. 2, each egg handling cup may be associated with asingle membrane. This makes it possible to analyze the VOCs emitted byeach individual egg. By incorporating a tracking mechanism into thesystem, the position of each egg can be matched with the membrane beinganalyzed. Once the analysis is complete, the eggs may be separated, byreference to the tracking system, on the basis of gender and/orfertility status.

An exemplary system and method of analyzing and sorting eggs is depictedin FIG. 3. In this embodiment, a conveyor has two reading trays (310 and315) to either side, in order to enhance throughput. In an embodiment,each reading tray includes one or more electromagnetic radiationtransmitters and detectors. Each reading tray may include a plurality ofholes (e.g., one for each egg handling cup. The holes act as apassageway through which electromagnetic radiation (e.g., THz or IRlight) can travel. In one embodiment, the egg handling cups of thevacuum egg handler, described in FIG. 2, are detachable from the vacuumhandling apparatus. For example, in an embodiment, a push-activatedlatching system may be used to mechanically detach and reattach the egghandling cups.

The system shown in FIGS. 2, 3, and 4 may be used to determine thegender and/or fertility status of avian eggs. Initially, eggs 304 may betransferred to conveyor 300 via carrier 302. Carrier 302 is part of thevacuum handling equipment. Carrier 302 includes a sub-manifold 240 (seeFIG. 4), which can be used to apply a vacuum to hold the eggs aspreviously described above relative to FIG. 2. The vacuum holding theeggs in place also causes VOCs to be transferred to the membranes, wherethe VOCs are retained. The vacuum is released and the eggs are placed ina crate 320 on the conveyor.

Carrier 302 may be coupled to a control system that controls vacuum tosub-manifolds 240. Carrier 302 is operable to selectively hold andrelease egg handling cups 220 and eggs, and to move egg handling cups220 relative to the conveyor and/or reading trays. Carrier 302 may bepositioned by way of motorized rail system, robotic arm, or othersuitable positioning mechanism. In some embodiments, carrier 302 movesegg handling cups 220 from the conveyor to one of the reading trays and,after reading, returns the egg handling cups to the conveyor.

To read the membranes, the sub-manifold 240 and empty egg handling cups220 are moved (e.g., via carrier 302) over to the first reading tray310. Sub-manifold 240 is lowered, and egg handling cups 220 are releasedto their appropriate spots on the first reading tray 310. The egghandling cups 220 are aligned with passageways 330 (show in tray 315).

Electromagnetic radiation transmitters and detectors (e.g., terahertz orinfrared electromagnetic radiation transmitters and detectors) are nowused to read the detached handling cups 220. FIG. 4 depicts anembodiment of a reading system 400. Reading system 400 includes aplurality of transmitters 410, a plurality of detectors 420, and amovable frame 430 which holds the transmitters and detectors inalignment with each other. Transmitters 410, detectors 420, and amovable frame 430 may be included in reading device 434. In theembodiment depicted, the movable frame 430 may be a sliding arm guidedby a track, wheels, or some other device that allows the frame to bemoved along a reading tray 310. In an embodiment, frame 430 includes atleast enough transmitters and detectors to read a single row ofmembranes disposed in the egg holding cups. In alternate embodiments,the frame may hold less than a row worth of transmitters and detectors(e.g., a single transmitter/detector pair). In such an embodiment, themovement mechanism of the frame may allow the frame to be moved in sucha way that each of the membranes in egg holding cups that were placed onreading tray 310 can be read. During use, the transmitters and detectorsare operated to read each of the membranes disposed in the egg holdingcups. A computer system 440 is coupled to the detectors to record thespectral information collected by the detectors. The spectralinformation is compared to spectral information collected in database445, which may be remotely located. This information is used todetermine the gender and/or fertility status of the eggs. Computersystem 440 and database 445 may be located at the facility, or at adifferent location (and remotely connected via, for example, a network).In one embodiment, computation relating to determination of fertilityand gender and/or storage of data relating to egg spectra areaccomplished remotely (e.g., through cloud services via a communicationnetwork).

The computer also includes tracking software that maps the obtainedspectral information to the location of the egg on the tray. Thetracking software may use bar codes or RFID on the trays (and/or palletson which trays are held) or the egg handling cups to track the positionof the eggs to the egg handling cup that captured the VOCs. After theanalysis is complete, the computer will send information to the vacuumegg handling apparatus to indicate which eggs need to be removed fromthe conveyor based on the gender and/or the fertility status. Sortingmay be accomplished by removing eggs from the vacuum egg handling devicebefore the eggs are transferred to crate 320. Alternatively, the eggsmay be sorted by placing the eggs on crate 320, then using further egghandling equipment downstream from the analysis component to separatethe eggs based on gender and/or fertility status.

While reading tray 310 is being scanned, carrier 302 may movesub-manifolds 240 to reading tray 315, wherein egg handling cups thathave been previously read can be reattached to the sub-manifolds. In oneembodiment, reading tray 315 may be used to clean the membranes from thepreviously read egg handling cups. In one embodiment, after all of theegg handling cups are read, a fluid stream (e.g., air or an inert gassuch as nitrogen) is passed through the passageways of reading tray 315(or reading tray 310) that removes VOCs from the membranes.Alternatively, an electrical system may be incorporated in the readingtrays so that an electrical current can be passed into the membrane toremove the VOCs. A combination of air or an inert gas and electricalcurrent may also be used. Alternatively, membranes may be discarded andreplaced with new, clean ones. In another embodiment, as the vacuumhandler lifts the egg handling cups, the vacuum handler may blow air oran inert gas through the membranes (e.g., by reversing the vacuummotor), incite an electric current in the membrane, or use a combinationof both methods to remove VOCs from the membrane.

After the sub-manifolds have been reloaded with the egg handling cups,the vacuum egg handling system uses the egg handling cups to obtain anew collection of eggs and place them on the conveyer. In an embodiment,the cleaning of membranes at reading tray 315 and collection of eggs iscompleted at about the same time that reading the membranes at readingtray 310 is completed. The cycle set forth above is repeated for eachcollection of eggs obtained, allowing more than 100 eggs to be sampledand tested within minutes.

Sample Chamber System

In another embodiment, a system for determining the gender and/orfertility status of avian eggs includes a sampling apparatus that can beused to collect volatile organic compounds (VOCs) from an intact egg. Inone embodiment, the sampling apparatus includes a vacuum source and asample chamber capable of holding volatile organic compounds emitted byan intact egg.

During use, a sampling apparatus applies a vacuum from the vacuum sourceto the gas proximate to the avian egg and directs the gas captured fromthe vicinity of the egg into the sample chamber. In an embodiment, thesampling apparatus includes a gas collection device which is placedproximate to the egg, as previously discussed.

In one embodiment, a vacuum egg handling system, such as shown in FIG.1, may be modified to include a plurality of sample chambers. FIG. 5depicts a schematic diagram of a sub-manifold 240 of a vacuum egghandling system that is modified to include a sample chamber 510 tocapture VOCs from eggs that are disposed in egg handling cups 220. Inthis embodiment, egg handling cups 220 act as the gas collection device.Gas in the vicinity of the egg is pulled through the egg handling cups220 into the sample chamber 510. Sample chamber 510 may include a topvalve 512 and a bottom valve 514. During use, a vacuum may be applied tosample chamber 510 from sub-manifold 240. While a vacuum is applied tosample chamber 510, bottom valve 514 is closed. Once an appropriatevacuum is obtained, top valve 512 is closed, creating a vacuum withinsample chamber 510. The egg handling cup 220, is positioned on the eggs,and bottom valve 514 is opened. The vacuum in sample chamber 510 pullsthe egg into the egg handling cup and draws any VOCs being released fromthe egg into the sampling chamber. After a predetermined time, or oncethe egg has been moved into position over a crate, bottom valve 514 isopened, breaking the vacuum to the egg as VOCs are drawn into thechamber, allowing the egg to drop into a crate, at which point valve 514is closed, sealing the collected VOCs in the sample chamber.

An alternate embodiment of a sub-manifold 740 of a vacuum egg handlingsystem is depicted in FIG. 7. The sub-manifold is modified to include asample chamber 750 which can be used to analyze VOCs captured from eggsthat are disposed in egg handling cups 720. In this embodiment, egghandling cups 720, in conjunction with sample chamber 750 act as a gascollection device. Each of the egg handling cups 720 includes a topvalve 712 and a bottom valve 714 which are used to define a collectionspace 710. Controller 760 operates the valves and the vacuum motor insequences that allow VOCs from each egg disposed in the egg handing cupsto be individually analyzed and associated with the egg.

During initial use, a vacuum may be created in collection space 710though sub-manifold 740. The vacuum may be applied by running vacuummotor (M) so that a vacuum is created in sample chamber 750. Samplechamber 750 includes two valves, 752 and 754. Valve 752 allows gases toenter sample chamber 750 from sub-manifold 740. Valve 754 controlsaccess of sample chamber 750 to vacuum motor (M). Initially, valves 752and 754 are opened and the vacuum motor is operated to create a vacuumin sample camber 750 and sub-manifold 740. This also has the effect ofpurging both sub-manifold 740 and the sample chamber 750 of any VOCsobtained from previous tests. In an optional embodiment, a purging gas742 may be introduced into sub-manifold 740 during evacuation of thesub-manifold and sample chamber. After a sufficient amount of purginggas is passed through the system, the purging gas is stopped and thesystem placed under a vacuum as discussed above. Top valves 712 are alsoopened, allowing a vacuum to be created in each collection space 710.Once a vacuum has been established throughout the system valves 754 and752 are closed, creating a sealed vacuum in the sample chamber, and topvalves 712 are closed creating a vacuum in each collection space 710.

During the next phase of use, egg handling cups 720, are positioned oneggs, and bottom valve 714 is opened. The vacuum in collection space 710pulls the egg into the egg handling cup and draws any VOCs beingreleased from the egg into the collection space.

The collected gas in each collection space is transferred to samplechamber 750 for detection of VOCs. In an embodiment, after VOCs havebeen collected in a collection space for a sufficient amount of time,the collected gas is transferred to sample chamber 750 by closing bottomvalve 714, opening the associated top valves 712, and opening samplechamber valve 752. Sample chamber 750 is at a lower vacuum, at thistime, than the rest of the system and therefore draws the collected gasinto the sample chamber 750 for analysis. Once the collected gas isdrawn into sample chamber 750, sample chamber valve 752 may be closed toretain the collected gas in the sample chamber. The gas in the samplechamber can be analyzed using techniques set forth herein to determinethe VOC content of the gas. The VOC content of the gas is then used todetermine the gender and/or fertility status of the egg in the positonassociated with the collected gas. Controller 760 keeps track of theposition of the egg handling cup associated with the open top valve 712and determines where the egg should be placed to allow sorting of theegg based on gender and/or fertility of the egg.

After detection of VOCs is complete, sample chamber 750 and sub-manifold740 are purged of the collected gas already present in the system beforeanalyzing another egg. To purge system vacuum motor (M) is run andsample chamber valves 754 and 752 are opened allowing gas from samplechamber a 750 and the sub-manifold 740 to be pulled out of the system.During cleaning of the system a purging gas 742 may be introduced intosub-manifold 740 during evacuation of the sub-manifold and samplechamber. Once the system is purged, the above process can be repeated onother eggs attached to the sub-manifold. In this way, each egg can beidentified for subsequent sorting.

An alternate embodiment of the system of FIG. 7 is depicted in FIG. 8.FIG. 8 is similar in many aspects to the system of FIG. 7, but differsin that sample chamber 750 includes a membrane 756 which is used tocapture VOCs in the collected gas. In FIG. 8, during initial use, avacuum may be created in collection space 710 though sub-manifold 740.The vacuum may be applied by running vacuum motor (M) so that a vacuumis created in sample chamber 750. Sample chamber 750 includes twovalves, 752 and 754. Valve 752 allows gases to enter sample chamber 750from sub-manifold 740. Valve 754 controls access of sample chamber 750to vacuum motor (M). Initially, valves 752 and 754 are opened and thevacuum motor operated to create a vacuum in sample camber 750 andsub-manifold 740. This also has the effect of purging both sub-manifold740 and the sample chamber 750 of any VOCs obtained from previous tests.Top valves 712 are also opened, allowing a vacuum to be created in eachcollection space 710. Once a vacuum has been established throughout thesystem top valves 712 are closed creating a vacuum in each collectionspace 710.

During the next phase of use, egg handling cups 720, are positioned oneggs, and bottom valve 714 is opened. The vacuum in collection space 710pulls the egg into the egg handling cup and draws any VOCs beingreleased from the egg into the collection space.

The collected gas in each collection space is transferred to samplechamber 750 for detection of VOCs. In contrast to the system depicted inFIG. 7, transfer of the collected gas is assisted by use of a membrane.In an embodiment, after VOCs have been collected in a collection spacefor a sufficient amount of time, the collected gas is transferred tosample chamber 750 by turning vacuum motor on (if the motor was turnedoff), opening sample chamber valves 754 and 752, and opening the topvalve 712 and bottom valve 714 associated with the egg being examined.The vacuum created by vacuum motor (M) draws the collected gas fromcollection space 710 into sample chamber 750 and through membrane 756.As the collected gas is drawn across the membrane, VOCs emitted by theegg are captured by the membrane. Once a sufficient amount of gasassociated with the egg is collected, sample chamber valves 752 and 754may be closed to movement of gas through membrane 756. The membrane inthe sample chamber can be analyzed using techniques set forth herein todetermine the VOC content of the gas. The VOC content of the gas is thenused to determine the gender and/or fertility status of the egg in thepositon associated with the collected gas. Controller 760 keeps track ofthe position of the egg handling cup associated with the open top valve712 and determines where the egg should be placed to allow sorting ofthe egg based on gender and/or fertility of the egg.

After analysis of membrane 756 is complete, the membrane may be cleanedor replaced as discussed above for other membrane systems. To preparefor the next reading, sample chamber 750 and sub-manifold 740 are purgedof any collected gas already present in the system before analyzinganother egg. To purge system vacuum motor (M) is run and sample chambervalves 754 and 752 are opened allowing gas from sample chamber a 750 andthe sub-manifold 740 to be pulled out of the system. During cleaning ofthe system a purging gas 742 may be introduced into sub-manifold 740during evacuation of the sub-manifold and sample chamber. Once thesystem is purged, the above process can be repeated on other eggsattached to the sub-manifold. In this way, each egg can be identifiedfor subsequent sorting.

Using these systems, each egg handling cup may be associated with asingle sample chamber. This makes it possible to analyze the VOCsemitted by each individual egg. By incorporating a tracking mechanisminto the system, the position of each egg can be matched with the samplechamber being analyzed. Once the analysis is complete, the eggs may beseparated, by reference to the tracking system, on the basis of genderand/or fertility status.

The system of FIGS. 3 and 4 may be used to analyze the VOCs collectedfrom the eggs, in a similar manner to the method used to analyze themembrane captured VOCs. In an embodiment, the egg handling cups of thevacuum egg handler, described in FIG. 5, are detachable from the vacuumhandling apparatus. The sub-manifold 240 and now empty egg handling cupsare moved over to the first reading tray 310. Sub-manifold 240 islowered and the egg handling cups are released to their appropriatespots on the first reading tray 310. The egg handling cups 220 arealigned with passageways 330. Electromagnetic radiation transmitters anddetectors (e.g., terahertz, microwave or infrared electromagneticradiation transmitters and detectors) are now used to read the samplechambers in the detached handling cups 220 in the manner describedabove. After the analysis is complete, the computer will sendinformation to the vacuum egg handling apparatus to indicate which eggsneed to be removed from the conveyor.

While one reading tray is being scanned, the vacuum handling machine maymove the sub-manifolds to a second reading tray, where egg handling cupsthat have been previously read can be reattached to the sub-manifolds,effectively doubling the rate at which eggs can be read in the system.In one embodiment, each reading tray may be used to clean VOCs from thesample chambers. In one embodiment, after all of the egg handling cupsare read, a stream of gas is passed through the passageways of a readingtray creating a stream of gas that removes VOCs from the samplechambers. Alternatively, as the vacuum handler lifts the egg handlingcups, the vacuum handler may blow air through the sample chambers (e.g.,by reversing the vacuum motor) to remove the VOCs from the samplechamber. During cleaning of the sample chamber, both valves 512 and 514are opened.

After the sub-manifolds have been reloaded with the egg handling cups,the vacuum egg handling system uses the egg handling cups to obtain anew collection of eggs and place them on the conveyer. The cycle setforth above is repeated for each collection of eggs obtained, allowingmore than 100 eggs to be sampled and tested within minutes.

Although in some embodiments described herein, a vacuum system was usedto hold and/or collect gas, a system may, in various embodiments,collect and/or capture gases emitted from the egg without application ofa vacuum.

Although in some embodiments described herein, gas collection devices(e.g., egg cups) are moved away from the egg, a system may, in variousembodiments, measure air or other gases collected near the egg withoutmoving the gas collection devices away from the egg.

In some embodiments, eggs may be read one after another (e.g., eggsarranged in a line sequentially on a conveyor belt system), rather thanthrough a batch process as described in FIGS. 1-3.

In Ovo Detection System

In an alternate embodiment, the VOC content of the air cell present inavian eggs may be directly analyzed using electromagnetic radiation thatis substantially transparent to the egg shell (i.e., in ovo VOCdetection). For example, both terahertz and microwave electromagneticradiation may be used for in ovo detection. In an embodiment, theappropriate transmitter and detector are placed in fixed positions withrespect to the air cell. A plurality of pairs of transmitters anddetectors can be used to analyze the VOC content of the air cell bymoving eggs through the optical pathway between the transmitter anddetector pair. The detected absorption spectrum is analyzed as discussedabove, and a determination between gender and/or fertility status may bemade. The analysis spectra may be generated using absorbance,transmittance, or reflectance through the egg shell.

In a first embodiment, eggs are positioned with their major axisvertically oriented in normal egg-crate-like storage containers, withthe air cell positioned at the top of the egg. Electromagnetic radiationtransmitters and detectors may be placed in optical communication withthe air cell of the eggs. In a second embodiment, the eggs arepositioned with their major axis vertically oriented, but with the aircell positioned at the bottom of the egg (closer to the ground). Aspecialized crate allowing line-of-sight access to the area of the eggcontaining the air cell is used to enable the electromagnetic detectionsystem to analyze the air cell content. As noted previously, the eggorientation can be either way.

In one embodiment, an egg handling system, such as shown in FIG. 1, maybe modified to include an in ovo detection system. FIG. 6 depicts aschematic diagram of an in ovo detection system. In this embodiment, acrate 320 of an egg handling system is modified to include one or morewindows 610 which will allow preselected electromagnetic radiation(e.g., terahertz, microwave or infrared radiation) to pass through thewindow and into the egg. In an embodiment, crate 320 may include aplurality of egg holding cups, which are shaped such that the egg isnaturally positioned with the air cell of the egg in optical alignmentwith the one or more windows. For example, the air cell of most eggstends to be positioned at the wider side of the egg. The crate may beshaped such that the bottom of the egg holding cups is wider than thetop to encourage proper alignment of the air cell with the window.

Each egg handling cup may be associated with a single individual egg. Byincorporating a tracking mechanism into the system, the position of eachegg can be matched with the egg handling cup holding the egg. Once theanalysis is complete, the eggs may be separated, by reference to thetracking system, on the basis of gender and/or fertility status.

In an embodiment, the one or more pairs of electromagnetic radiationtransmitters 620 and detectors 630 (e.g., terahertz, microwave orinfrared electromagnetic radiation transmitters and detectors) are nowused to read the air cell. In an embodiment, the transmitters anddetectors may be coupled to a movable frame (not shown) which holds thetransmitters and detectors in alignment with each other. The movableframe may be a sliding arm guided by a track, wheels, or some otherdevice that allows the frame to be moved along the crate. In anembodiment, the frame includes at least enough transmitters anddetectors to read a single row of eggs disposed in the egg holding cups.In an alternate embodiment, the movement mechanism of the frame mayallow the frame to be moved in such a way, e.g. to any point along a twodimensional plane, that each of the air cells of eggs in the egg holdingcups can be individually read. During use, the transmitters anddetectors are operated to read each of the air cells disposed in the eggholding cups. A computer system is coupled to the detectors to recordthe spectral information collected by the detectors. The spectralinformation is compared to spectral information collected in database.As noted previously, a computer system and database may be located atthe facility, or at a different location (and remotely connected via,for example, a network). In one embodiment, computations and/or storagefor testing and/or egg management are accomplished through cloudservices via a communication network. This information is used todetermine the gender and/or fertility status of the eggs. After theanalysis is complete, the computer will send information to the vacuumegg handling apparatus to indicate which eggs need to be removed fromthe conveyor.

While the system is depicted as reading through the “bottom” of thecrates, it should be understood that, in an alternate embodiment, in ovodetection can be carried out by analyzing the exposed (that is, “top” or“upper”) portion of the eggs disposed in the crate. In such anembodiment, it is necessary to place the egg in the crate with the aircell portion of the egg exposed so that a reading can be made withoutthe need for a window.

In this patent, certain U.S. patents, U.S. patent applications, andother materials (e.g., articles) have been incorporated by reference.The text of such U.S. patents, U.S. patent applications, and othermaterials is, however, only incorporated by reference to the extent thatno conflict exists between such text and the other statements anddrawings set forth herein. In the event of such conflict, then any suchconflicting text in such incorporated by reference U.S. patents, U.S.patent applications, and other materials is specifically notincorporated by reference in this patent.

Further modifications and alternative embodiments of various aspects ofthe invention will be apparent to those skilled in the art in view ofthis description. Accordingly, this description is to be construed asillustrative only and is for the purpose of teaching those skilled inthe art the general manner of carrying out the invention. It is to beunderstood that the forms of the invention shown and described hereinare to be taken as examples of embodiments. Elements and materials maybe substituted for those illustrated and described herein, parts andprocesses may be reversed, and certain features of the invention may beutilized independently, all as would be apparent to one skilled in theart after having the benefit of this description of the invention.Changes may be made in the elements described herein without departingfrom the spirit and scope of the invention as described in the followingclaims.

1-54. (canceled)
 55. A system for determining the gender and/orfertility status of avian eggs comprising: one or more carriers forreceiving one or more avian eggs; one or more transmitters capable ofemitting preselected electromagnetic radiation, wherein the one or moretransmitters direct the preselected electromagnetic radiation into theone or more avian eggs; and one or more detectors capable of detectingthe preselected electromagnetic radiation that passes through the one ormore avian eggs.
 56. The system of claim 55, wherein the electromagneticradiation is in the terahertz frequency range.
 57. The system of claim55, wherein the carrier is incorporated into an egg handling apparatus.58. The system of claim 57, wherein the egg handling apparatus comprisesat least one transmitter and at least one detector, wherein each of theat least one transmitter and at least one detector are capable ofsubstantially simultaneously transmitting the preselectedelectromagnetic radiation into a plurality of eggs.
 59. The system ofclaim 57, wherein the egg handling apparatus comprises a conveyancesystem for transporting eggs along a conveyor belt, wherein the carrieris coupled to the conveyor belt, and wherein at least one transmitterand at least one detector are positioned such that the transported eggsare brought into the optical pathway between the transmitters and thedetectors.
 60. The system of claim 55, wherein the carrier comprises oneor more windows positioned proximate to the location of avian eggs whendisposed in the carrier, wherein the one or more windows allow thepreselected electromagnetic radiation to pass through the window, andwherein the transmitter is optically aligned with the window such thatthe preselected electromagnetic radiation emitted by the transmitterpasses through the window and into the egg.
 61. The system of claim 60,wherein the carrier is movable such that the one or more windows can bemoved into the optical pathway between the transmitter and the detector.62. The system of claim 60, wherein the transmitter and/or the detectorare movable such that the one or more windows can be positioned into theoptical pathway between the transmitter and the detector.
 63. The systemof claim 60, wherein the carrier comprises one or more egg shaped cupsfor holding the avian eggs in a position such that the air sac of theegg is positioned in the window of the carrier.
 64. The system of claim60, wherein the carrier is in communication with a vacuum source, a gascollection device coupled to the vacuum source and a membrane positionedin the passageway coupling the vacuum source to the gas collectiondevice, wherein the membrane is capable of capturing volatile organiccompounds, wherein the sampling apparatus applies a vacuum from thevacuum source to the gas proximate to the avian egg and directs the gascaptured from the vicinity of the egg toward the membrane; wherein themembrane is positionable within the electromagnetic radiation emitted bythe transmitter.
 65. A method of determining the gender and/or fertilitystatus of avian eggs using a system as described in claim 55, the methodcomprising: positioning the air sac of an egg in the optical pathwaybetween the transmitter and the detector; and applying electromagneticradiation to the air sac of the egg, wherein the electromagneticradiation is preselected to allow the quantity of one or more organiccompounds in the air sac of the egg to be determined.
 66. The method ofclaim 65, wherein the carrier is incorporated into an egg handlingapparatus; and wherein the egg handling apparatus comprises a pluralityof the transmitters and detectors, and wherein the method furthercomprises substantially simultaneously transmitting the preselectedelectromagnetic radiation into a plurality of eggs.
 67. The method ofclaim 65, wherein the electromagnetic radiation is in the terahertzfrequency range.
 68. The method of claim 66, wherein the egg handlingapparatus comprises a conveyance system for transporting eggs along aconveyor belt and a plurality of transmitters and detectors, and whereinthe method comprises moving the carrier along the conveyor belt suchthat the transported eggs are brought into the optical pathway betweenthe transmitters and the detectors.
 69. The method of claim 65, whereinthe carrier comprises one or more windows positioned proximate to thelocation of avian eggs when disposed in the carrier, wherein the one ormore windows allow the preselected electromagnetic radiation to passthrough the window, and wherein the method further comprises opticallyaligning the transmitter with the window such that the preselectedelectromagnetic radiation emitted by the transmitter passes through thewindow and into the egg.
 70. The method of claim 68, further comprisingmoving the carrier such that the one or more windows are moved into theoptical pathway between the transmitter and the detector.
 71. The methodof claim 65, further comprising moving the transmitter and/or thedetector such that the one or more windows are positioned in the opticalpathway between the transmitter and the detector.
 72. The method ofclaim 65, wherein the carrier is in communication with a vacuum source,a gas collection device coupled to the vacuum source and a membranepositioned in the passageway coupling the vacuum source to the gascollection device, wherein the membrane is capable of capturing volatileorganic compounds, wherein the sampling apparatus applies a vacuum fromthe vacuum source to the gas proximate to the avian egg and directs thegas captured from the vicinity of the egg toward the membrane; whereinthe membrane is positionable within the electromagnetic radiationemitted by the transmitter.